Donor age and gestational age influence on growth factor levels in human amniotic membrane

Abstract

Amniotic membrane (AM) is used as a biomaterial for reconstruction in ocular surface surgery. This study investigated the influence of interdonor variations and processing and preservation procedures applied to the AM on growth factors and protein levels. Samples of human AM from thirteen donors were analysed. Collected donor data were age, parity and gestational age. Total protein amount was measured in extracts of intact AM nonpreserved, lyophilized and cryopreserved, at -80°C and in liquid nitrogen. An enzyme-linked immunosorbent assay (ELISA) was used to assay growth factors protein levels for epidermal growth factor, basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), transforming growth factor beta1 (TFG-β(1) ) and nerve growth factor (NGF). Univariate and multivariate statistical analyses were used to study the influence of the preservation method applied and interdonor variations on growth factors levels. We detected important variations in growth factors and protein concentrations between samples from different donors. Total protein amount, bFGF, HGF, KGF and TGF-β(1) showed lower levels in samples from donors with higher gestational ages and donor ages, for all groups. The variability in the biochemical composition of AM from different donors is considerable, and it is related with donor factors as donor age and gestational age. As AM biochemical composition has a role in its therapeutic effects, these variations could affect the clinical results of amniotic membrane transplantation and must be taken into account in donor selection processes.